INFECTION.

INFECTIOUS PROCESS
Learning guide for the 2nd and 3rd year English media
students of the Faculty of Medicine and the Faculty
of Dentistry (Microbiology, virology and immunology)

ІНФЕКЦІЯ.
ІНФЕКЦІЙНИЙ ПРОЦЕС
Методичні вказівки з дисципліни
"Мікробіологія, вірусологія та імунологія"
для студентів II і III курсів медичного
та стоматологічного факультетів
з англійською мовою викладання

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 МІНІСТЕРСТВО ОХОРОНИ ЗДОРОВ’Я УКРАЇНИ
Харківський національний медичний університет

INFECTION.
INFECTIOUS PROCESS
Learning guide for the 2nd and 3rd year English media
students of the Faculty of Medicine and the Faculty
of Dentistry (Microbiology, virology and immunology)

ІНФЕКЦІЯ.
ІНФЕКЦІЙНИЙ ПРОЦЕС
Методичні вказівки з дисципліни
"Мікробіологія, вірусологія та імунологія"
для студентів II і III курсів медичного
та стоматологічного факультетів
з англійською мовою викладання
Затверджено вченою радою ХНМУ.
Протокол № 10 від 21.11.2019.

Харків
ХНМУ
2019

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 Infection. Infectious process : learning guide for the 2nd and 3rd year
English media students of the Faculty of Medicine and the Faculty of Dentistry
(Microbiology, virology and immunology) / N. I. Kovalenko, T. M. Zamaziy. –
Kharkiv : Kharkiv National Medical University, 2019. – 36 p.

Compilers N. I. Kovalenko,
T. M. Zamaziy

Learning guide is related to the program of Ministry of Health of Ukraine

and is recommended to students of medical and dentistry faculties of high
medical schools of III–IV level accreditation.
Learning guide includes sections of characteristics of infectious process
and types of infections, epidemiology and specifity of infectious diseases,
classification of bacteria due to pathogenicity, virulence factors of bacteria. The
most modern information on biological method of diagnosis of infectious
diseases is represented.

Інфекція. Інфекційний процес метод. вказівки з дисципліни:

"Мікробіологія, вірусологія та імунологія" для студентів II і IІІ курсів мед.
та стомат. фак-тів з англ. мовою викладання / упоряд. Н. І. Коваленко,
Т. М. Замазій. – Харків : ХНМУ, 2019. – 36 с.

Упорядники Н. І. Коваленко,
Т. М. Замазій.

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 Theme: Studies about an infection.
Experimental method of diagnostics of infectious diseases
Actuality of the theme
The infection (Latin of infection – to infect, pollute) is a form of relationship
which evolutionarily developed between pathogenic microbes and human in
certain environmental conditions.
Manifestations of an infection are various and also depend on properties of
a microorganism, state of a macroorganism and environmental conditions.
Infectious process is a collection of physiological protective and pathological
reactions that occur in a macroorganism in response to the action of pathogenic
microbes. It develops consistently and involves a complex of biochemical,
cytochemical, morphological changes that occur in the body.
Infectious disease is the extreme stage of the infectious process. Infectious
disease is clinical manifestation of an infectious process, accompanied by
characteristic signs or symptoms.
The microbes that cause infectious diseases are commonly called the
causative agents of infectious diseases. A human or animal organism that is in a
state of infection, that is, parasitization of a pathogen in it, is called infected,
while environmental objects that have been affected by pathogens should be
designated as contaminated with one or another pathogen. The susceptibility of
a macroorganism should be understood as the ability of a macroorganism to
respond to the introduction of microbes by the development of an infectious
process in its various manifestations - from carriage to infectious disease.
Epidemiology of infectious disease
The epidemic process is a continuous course of interconnected infectious
diseases caused by the presence of a source, a transmission mechanism of the
causative agent of infectious diseases, and a susceptible organism.
Participants of the epidemic process (Fig. 1):
– Source (reservoir) of infection
– The mechanism and route of transmission
– Susceptible organism
The source of infection is an abiotic object, a living organism, where the
pathogenic microbe lives, from which infection occurs. The source can be a
patient, a carrier, abiotic objects (water, food).
Classification by source of infection:
– sapronous infections are diseases whose main habitat and reproduction of
pathogens are environmental objects, from where they enter the human body
(diseases caused by legionella, Pseudomonas aeruginosa, etc.);
– anthroponous infections are diseases in which the only source of the
pathogen is a person (meningococcal infection, dysentery, cholera, diphtheria,
syphilis, hepatitis B, epidemic typhus, epidemic relapsing fever, etc.);
– zoonotic infections are diseases in which the only source of the pathogen
is animals (tularemia, brucellosis, rabies);

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 – zooanthroponic infections are diseases in which the source is an animal
and a sick person, including the corpses of the dead (plague, anthrax,
tuberculosis, rickettsioses).

Fig. 1. Participants of the epidemic process

The transmission mechanism is the process of removing the pathogen from
the host organism into the environment, its stay on environmental objects, the
introduction of the pathogen into a susceptible organism.
Transmission mechanism is a method for moving a pathogen from an
infected organism to a susceptible one.
It is carried out in 3 phases:
1. Excretion of the pathogen from the host.
2. Stay in the environment.
3. The introduction of the pathogen into a susceptible organism.
Factors of transmission are environmental objects that transfer microbes
from one organism to another.
Routes of transmission are a combination of factors in certain environmental
conditions, ensuring the pathogen from one macroorganism to another.
Entrance gates (portal of entry) are organs and tissues of the host organism
through which pathogenic microbes penetrate.
The entrance gate of infection can determine the clinical form of the disease. The
same pathogen microorganism, entering the macroorganism in various ways, causes
different clinical forms of the disease, as is the case with anthrax:
• cutaneous form is caused by the penetration of microorganisms into the
body through the skin;
• pulmonary – through the mucous membranes of the upper respiratory tract;
• intestinal – gastrointestinal tract.

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 The susceptibility of society is the presence of an immune layer in the
population against the corresponding pathogen.
The allocation of a particular transmission route of infectious diseases is
rather conditionaly. According to the mechanism of transmission infectious
diseases are divided into:
1. intestinal (fecal-oral);
2. respiratory;
3. infections of the blood (trasmissive);
4. infections of the skin and mucous membranes (contact). In is divided into:
• direct contact – from a source to a host, including sexually transmitted
diseases (for example HIV infection);
• indirect contact – through an intermediate object – it can be hands (for
wound infections, intestinal infections) or various objects, including medical
ones (for purulent-inflammatory diseases and parenteral hepatitis);
5. vertical (from mother to fetus).
Nosological form of the disease depends on the route of transmission of a
pathogenic microorganism:
• when ingested, streptococci cause tonsillitis;
• household contact - streptodermia (purulent-inflammatory disease of the skin).
Infectious diseases are characterized by infectiousness and can occur in the
form of epidemics - diseases registered in large territories (region, country) and
associated with the source of infection. Diseases spread to several countries and
continents are called pandemic (influenza, cholera). Infectious diseases that
occur in rare cases are called sporadic. Diseases prevalent only in certain areas
where there is a reservoir and vector for infection are called endemic
(tularemia, tick-borne encephalitis). They are usually caused by natural factors
that are favorable for the development of pathogens and reservoirs of the
pathogen. Natural infections exist naturally in wild animals and from time to
time cause outbreaks or epizootics. In case of accidental exposure to such area,
a person can become infected and get sick (plague, tularemia, tick-borne
encephalitis, rickettsiosis).
Depending on the route of penetration of the pathogen into the host,
exogenous and endogenous infections are distinguished.
Exogenous infections occur due to the entering of pathogens from the
environment. Infection can occur through damaged skin, mucous membranes of
the eyes, respiratory, urogenital tract, gastrointestinal tract. Even microtraumas,
insect bites, needle punctures can cause infection.
In endogenous (autoinfection) infection, pathogens are found in the body in
the obligate or transit flora. When the protective properties of the body are
weakened, they can be the cause of the disease (candidiasis).
Depending on the manifestations infections are manifest and inapparent.
Manifest – with typical and atypical manifestations of the disease. Typical –
all clinical signs appear, periods are expressed. Atypical – some characteristic

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signs are absent. For example: cholera without diarrhea, typhoid fever without
rash, etc. It is very difficult to diagnose this disease and only microbiological
methods of study are crucial.
This type of infection is characterized by a short stay of the microbe in a
macroorganism lasting up to 3 months, which in some cases goes into a
protracted form of the infection, lasting from 3 to 6 months. A protracted (or
subacute) form is characterized by an increase in the invasive period and the
period of convalescence and is often a transition from acute to chronic infection.
Inaparantic (latent, subclinical) – no external manifestations of the disease,
but there are immunological, morphological, typical changes for the corresponding
disease. The activator is exposed to the pressure of the protective forces. The
pathogen will stay in the patient's body for a long time without showing itself.
With the weakening the body (cooling, fatigue, fasting) latent infections can
turn into a typical disease with a pronounced clinical picture.
Diagnosis of inapparent forms of infection is possible only in the foci of infectious
diseases on the basis of specific laboratory methods (change in the rise of antibody
titers in dynamics, morphological studies, skin allergic tests, etc.).
This type on infections is characterized by a long stay of the microbe in a
macroorganism, or persistence (from lat. Persistentia - preservation of a
previous state, constancy). Persistence confirms the inability of society and the
macroorganism to cope with the microbe and the ability of the latter to survive
in the macroorganism. The mechanisms for the development of persistence are
diverse. An important role is played by: the formation of morphologically
modified or defective forms of microbes (L-forms of bacteria, cysts, defective
viral particles); the formation of drug resistance, the ability of microbes to
intracellular parasitization (pathogens of malaria and Leishmania, viruses,
chlamydia, etc.); blocking apoptosis of host cells; the presence of both
congenital and acquired immunodeficiencies, including under the influence of
microbes; development of immunological tolerance, as well as autoimmune and
allergic reactions in a macroorganism, etc. A distinctive feature of persistence
is that it develops against the background of acquired immunity, which is
ineffective, and also that it is a pathogenetic norm for a number of microbes
(viruses, rickettsia and chlamydia, mycobacteria, treponema, brucella, causative
agent of four-day malaria). Persistence may occur in several forms.
One form of infection that occurs without signs of the disease is carriage - a
condition of the body in which pathogens is present in the body and their release
into the external environment is not accompanied by clinical symptoms of the
disease and disorders by organs and tissues. It is formed more often after the
disease, when clinical recovery, but the pathogens continue to remain in the
body of the diseased and released into the environment (for example, carrying
typhoid, dysentery rods, diphtheria, cholera). In some cases, carriage develops
in healthy individuals who had contact with patients or even carriers of pathogens.

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 Acute carriage occurs when the pathogens persist in the body for 3 months,
chronic – longer (typhoid throughout life). Usually the carriage occurs in an
unacceptable organism against the background of immunity, which was formed
as a result of previously transferred disease or vaccinations. Bacterial carriers
contribute to the emergence and support the spread of infectious diseases in the
inter-epidemic period, when the spread of the pathogen due to natural causes
dies down (for example, when cold seasons occur with intestinal infections).
Latent infection (syn. dormant infection) is a peculiar form of carrier, in
which the microbe is present in the macroorganism for a long time, but is not
released into the environment. As a rule, latent infection is a natural stage of the
infectious process in diseases prone to a chronic course (brucellosis, syphilis,
herpetic infection, toxoplasmosis).
Chronic infection lasts more than 6 months and can occur in the form of a
continuous or relapsing form, characterized by a change in periods of
remissions and exacerbations, in which microbes are released into the
environment for many months and even years. Primary chronic diseases include
brucellosis, tuberculosis, leprosy, malaria and syphilis.
In virology, a slow virus infection has been identified as a separate group.
If the infection is caused by one type of pathogen, this infection is called
monoinfection. When infecting the body 2–3 different pathogens at the same
time (for example, diphtheria rods and streptococcus) mixed infection occurs.
A secondary infection is distinguished from mixed infections when an
underlying disease (such as influenza) is associated with an infection caused by
another pathogen (such as staphylococcus or streptococcus), as a result of
weakening the body's defenses. In people with immunodeficiency it is called
opportunistic infection.
Reinfection is called a condition when a new disease has occurred due to a
new infection with the same type of pathogen after recovery (gonorrhea). If the
disease has returned before recovery as a result of infection with the same
pathogen, it is superinfection (syphilis).
Relapse is the return of symptoms (typhoid fever, malaria) that occur
without re-infection by pathogens remaining in the body.
Depending on the localization of pathogens in the body of the patient there
distinguish a local infection, in which the microbes persist in one, certain place,
not spreading beyond its boundaries (eg, sore throat, furunculosis), and
generalized infection when the forces of aggression of microorganisms
overtake defense mechanism of host and pathogens from the local focus spread
throughout the body. A condition where the pathogens circulate for some time
in the blood, but do not multiply in it (for example, typhoid fever, brucellosis),
is called bacteremia, virusemia.
In the case when the pathogen is circulating in the blood for a long time,
accumulates and multiplies, sepsis or septicemia occurs (from lat. Sepsis - pus).
Such accumulation of microorganisms occurs in plague, anthrax. Sepsis is also

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caused by purulent cocci. The peculiarity of sepsis is that the clinical picture
does not depend on the type of pathogen.
The formation of purulent foci in various organs due to sepsis is called
septicopyemia. Circulation of the toxin in the blood is called toxinemia.
Participants in the infection process
Factors of the onset and outcome of the infectious process:
1. Quantitative and qualitative characteristics of microorganisms determine
the specificity of the infectious process.
2. The state of the macroorganism, its susceptibility to the microorganism
determines the form of manifestation, duration, severity and outcome of this
infectious process.
3. Environmental factors where the meeting of the microorganism with the
host has an indirect effect, reducing or increasing the susceptibility of the host
or the infectious dose and virulence of the pathogen.
Stages of the infectious process
The infectious process is one of the most dynamic forms of interaction
between microbes and a macroorganism that has developed during evolution.
The process proceeds with a constant change of cause and effect relationships.
Conventionally, it can be divided into several stages.
1. The penetration of the microorganism into the macroorganism, its
adaptation at the entrance gate and adhesion. The starting point of the infection
process is the introduction and adaptation of microbes (from late Lat. Adaptatio
- adaptation) at the site of the entrance gate of infection, as well as adhesion of
microbes to the cells of the macroorganism. Entrance gates are tissues and
organs through which microbes enter the body. In most cases, microbes enter
the microorganism through damaged skin and intact mucous membranes
permeable to microbes.
2. Colonization and formation of enzymes, toxins and other products during
the life and reproduction of microorganisms, which leads to disruption of
homeostasis due to local and generalized inflammation.
The second stage is colonization (from Lat. Colonia – settlement) –
horizontal colonization of the skin and mucous membranes at the site of the
entrance gate of infection. In the infectious process, the spread of microbes
occurs not only horizontally, on the surface of the cells, but also in the depths
of the cells and tissues of the macroorganism. The ability of microbes to
penetrate the cells of a macroorganism is called penetration. In this case, the
multiplication of microbes and the formation of new generations of the
pathogen under favorable conditions, as well as the release of metabolic
products of microbes, their enzymes and toxins and, in addition, the formation
of toxic decomposition products of macroorganism cells that have local or
long-term damaging effects on tissues and organs.
3. The third stage is dissemination (from the Latin. Disseminare – scatter,
spread) i.e., the spread of microbes beyond the primary focus of the

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introduction and colonization of microbes by the hematogenous pathway,
bronchogenic or perineural, along the nerve trunks, which leads to the
generalization of the infectious process (generalization is a transition from
general to particular, spreading throughout the macroorganism).
4. The fourth stage is the mobilization of the protective factors of the
macroorganism. In response to the penetration of microbes and their pathogenic
effect, the macroorganism mobilizes all initially non-specific and then specific
protective factors inherent in it, the action of which is aimed at neutralizing
both the microbes themselves and their toxins and at restoring impaired
homeostasis in the macroorganism.
5. The fifth stage is the end and outcome of the infectious process. In most
cases, sanitation of the macroorganism occurs (from the English. Sanative -,
healing) that is, the complete release of the macroorganism from the microbe
and the acquisition of a new quality by it - the formation of immunity. In some
cases, the infectious process ends in death. In those cases when equilibrium is
established between the microbe and the macroorganism, carrier state
formation takes place.
As a result of the action of many factors, the infectious process does not
always go through all its inherent stages and can end already in the early stages,
for example, proceeding in an abortive form of an infectious disease in
vaccinated or in persons who have previously undergone this disease. Another
example is the defeat of the mucous membranes of the urogenital tract in
gonorrhea without subsequent generalization of the infection, or colonization of
the mucous membranes in intestinal diseases without subsequent generalization
of the infection. In diphtheria, the infectious process is also limited by
adhesion, colonization and production of exotoxin (histotoxin). The penetration
of bacteria into the blood does not occur. If for extracellular parasites the
process is limited by adhesion and colonization, then for obligate and
facultative intracellular parasites, an important condition is their penetration
into the cell and subsequent intracellular multiplication.
The infectious process can occur at all levels of the organization of the
biological system of a macroorganism. In addition, each higher level includes
lower levels. First of all, the multi-level system of the infectious process
includes the body level or the infectious process itself, since infection is a
system of reactions that arise in a susceptible macroorganism. The subordinate
levels are the tissue-organ, cellular (the interaction of the microbial cell and the
host cell) and the subcellular or molecular level, which is based on the
competitive interaction of the biological molecules of the microbe and the
macroorganism. As a result of the interaction of the microbe with the
macroorganism, first of all, the cell suffers in which the microbe makes the
gaps through which it subsequently penetrates the macroorganism. The
interaction process itself occurs at the level of complementary structures of the
microbial macromolecules and the eukaryotic cells of the macroorganism. The

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processes taking place at the molecular and cellular levels, namely the
interaction of pathogenicity factors of microbes with cellular and humoral
factors of defense of a macroorganism, are reflected at the tissue-organ level.
Tissue differentiation of host cells determines the specificity of the infectious
process and plays a protective role, limiting the breeding zones of microbes.
Therefore, the main features of the pathogenesis of the infectious process are
formed at the tissue-organ level, reflected subsequently at the organismic level,
determining the manifestation of the infectious process. The features of the
pathogenesis and clinic of the infectious process at the organismic level are
reflected in the course of the epidemic process at the ecosystem level,
determining the nature and intensity of the implementation of a particular
mechanism of transmission of the pathogen. Thus, the infectious process is
characterized by a variety of complex interactions both at each level of the
system, and between these levels.
Pathogenicity and virulence of microorganisms
According to this property, microorganisms are divided into:
1. pathogenic;
2. opportunistic microorganisms occur in the environment and in the normal
microflora, safe for a healthy person, but can become pathogenic for persons
with immunodeficiency, when penetrating into organs where they are not
normally found (E. coli when penetrating the abdominal cavity in trauma or
during surgery causes peritonitis);
3. non-pathogenic.
Microorganisms that can cause disease in humans and animals are called
pathogenic, and their ability to cause disease is called pathogenicity (from the
Latin pathos – suffering, genos – birth). Pathogenicity is a genetically determined
species trait. Most pathogens are characterized by specificity that is the ability
of this type of microbe to cause a particular disease. For example, cholera is
caused by Vibrio cholera, gonorrhea – by gonococcus, dysentery – by Shigella
dysentery. However, the specificity of microorganisms is relative. So some
microorganisms (staphylococci and streptococci) can cause different diseases
and on the contrary the same diseases are caused by different pathogens. So
pneumonia can be caused by staphylococci, streptococci, Klebsiella and others
microorganisms.
Organotropy is the ability to affect certain tissues selectively (gonococcus
affects the epithelium of the genitals, the causative agent of dysentery -
enterocytes).
In order to cause an infectious process, pathogenic microbes must penetrate
the body in a certain critical infectious dose (pathogenic), i.e. in a minimum
dose, which causes persistent adhesion, colonization, penetration of a pathogen
into the tissue and the further development of the infectious process. For each
type of microbe, there is its own minimum infectious dose, i.e. the number of
individuals capable of causing the disease.

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 Different strains of the same species may have different pathogenic effects.
The degree of pathogenicity is called virulence.
Virulence (Lat. Virulentus – poisonous) is a sign not of a species, like
pathogenicity, but of a strain, i.e., it is inherent not to the whole species, but to
specific strains. Virulence can also be defined as the phenotypic manifestation
of the pathogenic genotype of microorganisms. As a quantitative sign, in
contrast to qualitative – pathogenicity, virulence has units of measure: it is
measured by the dose of microorganisms that cause a certain biological effect.
Strains are divided into: high, moderate, weak, avirulent.
Laboratory virulence is determined by the magnitude of the lethal (LD) and
infectious (ID) doses for experimental animals.
Lethal dose (LD) – the smallest number of pathogen or toxin, causing the
death of a specific number (%) of animals in a certain period of time.
• DCL (dosis certae letalis) – absolutely lethal dose – the minimum amount
of the pathogen that causes the death of 100 % of laboratory animals taken in
the experiment;
• DLM (dosis letalis minima) – the minimum lethal dose – the minimum
amount of the pathogen, causing the death of 95% of laboratory animals taken
in the experiment;
• LD50 – the minimum amount of the causative agent causing death of 50 %
of laboratory animals taken in the experiment (used to measure virulence most
often). At the same time, the type of laboratory animal at which this dose was
determined is always indicated, since the sensitivity of different types of
laboratory animals to various microorganisms is different.
In order to cause an infectious process, pathogenic microbes must penetrate
the body in a certain critical infectious dose (pathogenic), i.e. in a minimum
dose, which causes persistent adhesion, colonization, penetration of a pathogen
into the tissue and the further development of the infectious process. For each
type of microbe, there is its own minimum infectious dose, i.e. the number of
individuals capable of causing the disease.
Infectious dose (ID) – the minimum number of microorganisms that can
cause disease in a certain number (%) of experimental animals.
ID100 is the minimum number of living microbes that causes the
development of an infectious disease in 100 % of infected experimental animals
taken in the experiment.
ID50 – 50 % incidence, etc.
As a phenotypic manifestation of the genotype, virulence is subject to
changes both in the direction of decrease and in the direction of increase under
the influence of physical, chemical and biological factors. A decrease in
virulence (attenuation) can occur with prolonged cultivation of bacteria on an
artificial nutrient medium or as a result of prolonged passage of microbes
through the body of unreceptive animals. The complete loss of virulence is
associated with a change in genotype. An increase in virulence is observed, on

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the contrary, with the passage of microbes through the body of highly susceptible
animals, with lysogenesis, as well as due to mutations and recombinations.
These features of changes in virulence are taken into account when receiving
vaccine strains of microbes.
With a maximum decrease in virulence, pathogenic microorganisms can
become avirulent, i.e., non-virulent. But virulent microorganisms are always
pathogenic.
Virulence factors
The virulence of microorganisms is determined by three main properties of
the pathogen: infectivity, invasiveness, toxicity (Fig. 2).

Fig. 2. Bacterial mechanism of pathogenicity

Infectivity, invasiveness and toxicity are not related and are differently
detected in various pathogens. The pathogen of diphtheria, tetanus, botulism
more pronounced toxigenic properties. The plague agent is invasive. It does not
only suppress the body's defenses, it also multiplies so rapidly that the immune
system does not have time to react.
Infectiousness is the ability to infect. It is conditioned by its ability to adhere
(stick), colonize (multiply). Adhesion and colonization is trigger mechanism of
disease development. Microorganisms and toxins realize their pathogenic and
toxic properties when they bind to target cell receptors.
Adhesion is the ability to adsorb on cells of host organism that are sensitive
to a particular microorganism. It is caused, on the one hand, by the surface
structures of the microbial cell (pili, etc.), on the other, by the presence of
receptors of the cell of a macroorganism capable of interacting with the
microbial cell.
Colonization can be on the surface of cells to which germs adhere (Vibrio
cholera adheres to enterocytes), or inside cells into which adherent germs
penetrate (eg, S. dysentery multiplys in cells of the colon).

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 Invasiveness is the ability to penetrate the tissues, overcoming the body's
defenses. Invasiveness is related to the ability of germs to produce enzymes that
disrupt (increase) the permeability of connective and other tissues. For the
implementation of colonization and invasion, many bacteria secrete enzymes of
aggression and defense:
hyaluronidase (a spread factor), which destroys the hyaluronic acid of the
connective tissue and thereby promotes the penetration of germs into the tissue.
Neuraminidase, which cleaves neuraminic acid from glycoproteins,
glycolipids, polysaccharides that are part of different tissues, thus increasing
their permeability.
Fibrinolysin dissolves a clot of fibrin, which is formed during inflammation,
preventing the penetration of the microorganism into the tissues.
Collagenase ferments collagen of muscle fibers, causing their melting.
Lecithinase C ferments the lecithin of the membranes of muscle fibers, red
blood cells, and other cells.
Coagulase coagulates blood plasma, promotes the formation of fibrin
barriers.
Deoxyribonuclease (DNase) causes depolymerization of DNA.
Proteases destroy proteins (including IgG and others).
Substances with antifagocytic activity. Inhibition of phagocytosis is carried
out by bacterial capsules. The substances that make up the capsules of different
microorganisms are different, and their functions are also different. Thus, the
capsular polypeptide of the Bacillus anthracis protects it from capture by
phagocytes; polysaccharide of Pseudomonas aeruginosa suppresses both capture
and intracellular digestion of bacteria. Other capsular polysaccharides and
polypeptides of microorganisms: K- and Vi-Ag microcapsules of enterobacteria,
M-protein of β-hemolytic streptococci A, A-protein of staphylococci.
In addition to these factors, germs protect against phagocytosis by some
enzymes. For example, staphylococcus coagulase promotes plasma coagulation,
leading to the formation of a protective "cover" around the microbial cell;
fibrinolysin dissolves fibrin, contributing to the spread of germs.
The formation by microorganisms substances capable of withstanding
intracellular digestion; preventing the fusion of phagosome and lysosomes;
capable of causing lysis of phagocytes (leukocidins).
Antigenic mimicry is the similarity of the Ag-determinants in the
microorganism and the host macroorganism, as a result, the microorganism is
not recognized by the immune system, which contributes to its persistence in
the macroorganism.
Bacteria produce two types of toxins – exotoxins and endotoxins (table).

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 Table
Properties of bacterial exotoxins and endotoxins
Property Endotoxin Exotoxin
Source Gram-negative bacteria Gram-positive bacteria
Chemical nature Lipopolysaccharide Protein
Relationship to cell Part of outer membrane Extracellular, diffusible
Denatured by boiling No Usually
Antigenicity Weakly Highly
Form toxoid No Yes
Potency Relatively low (> 100 ug) Relatively high (1 ug)
Specificity Low degree High degree
Enzymatic activity No Usually
Pyrogenicity Yes Occasionally

Exotoxins are the products of the metabolism of germs released into the
environment. They have a protein origin, which causes their low resistance to
external actions. The exceptions are the neurotoxin of the botulinum rods, the
enterotoxins of staphylococcus, Vibrio cholera, which withstand short boiling.
Exotoxins are proteins of a bifunctional structure – the transport group
interacts with cell receptors, and the toxic group (activator) penetrates the cell
and blocks metabolic processes.
Exotoxins are mainly synthesized by Gram-positive bacteria (diphtheria,
tetanus, botulism, gas gangrene, S. aureus, S. pyogenes) and some Gram-
negative bacteria (V. cholera, P. aeruginosa, E. coli, S. dysentery).
According to the mechanism of action on the cells of a macroorganism,
bacterial toxins are divided into several types, although this division is quite
arbitrary and some toxins can be assigned to several types at once:
• Type 1 – membranotoxins (hemolysins, leukocidins);
• Type 2 – functional blockers, or neurotoxins (thetanospasmin, botulinum
toxin) - block the transmission of nerve impulses in synapses (in the cells of the
spinal cord and brain);
• Type 3 – thermostable and thermolabile enterotoxins – activate cell
adenylate cyclase, which leads to disruption of enterosorption and the
development of diarrhea syndrome. Such toxins produce V. cholera (cholerogen),
enterotoxigenic E. coli;
• Type 4 – cytotoxins – toxins that block protein synthesis at the subcellular
level (enterotoxin of S. aureus, anthrax bacilli, and B. pertussis); here also
include anti-elongators – preventing elongation (growth) or translocation, that

14
is, the movement of i-RNA along the ribosome, and thereby blocking protein
synthesis (diphtheria histotoxin, P. aeruginosa toxin);
• Type 5 – exfoliatins formed by certain strains of S. aureus and
erythrogenins produced by group A pyrogenic streptococcus (S. pyogenes).
They affect the interaction of cells with each other and with extracellular
substances and completely determine the clinical picture of infection (in the
first case pemphigus of newborns, in the second – scarlet fever).
Many bacteria form not one, but several protein toxins, which have different
effects - neurotoxic, cytotoxic, hemolytic: staphylococcus, streptococcus.
The ability of microorganisms to form protein toxins must also be
considered when conducting microbiological diagnostics. It should be
remembered that all pathogenic strains of this species can produce only one
type of toxin according to the antigenic structure and mechanism of action
(C. diphtheriae, C. tetani), different in antigenic structure, but are identical in
mechanism of action (C. botulinum). At the same time, some bacteria can
simultaneously form both protein exotoxins and endotoxins: E. coli, V. cholera.
Exotoxins are characterized by high toxicity and expressed specificity –
organotropicity. Each type of toxin affects certain organs or tissues. For
example, tetanus toxin affects the motor neurons of the spinal cord, and
diphtheria toxin affects the muscles of the heart and adrenal glands, botulinum
toxin acts on the end of motor nerves.
The synthesis of protein toxins is encoded by genes localized on the
chromosome and linked to genes involved in spore formation or part of the
prophage, as well as genes localized in plasmids. These are tox+genes
responsible for toxigenicity. The activity of tox+genes is controlled by
microbial cell repressor proteins. The initial function of these genes in
saprophytes was the synthesis of structural phage proteins, spore membrane
components, or the synthesis of enzymes necessary for the absorption of amino
acids. As the parasitic lifestyle consolidates, these specialized adaptive
enzymes have turned into poisons – protein toxins.
Endotoxins are a lipopolysaccharidoprotein complex that is closely related
to the cell of the microorganism and is a part of the cell wall mainly of Gram-
negative bacteria, released after cell destruction (typhoid, dysentery, cholera,
whooping cough). They are nonspecific. The entire LPS molecule is responsible for
the manifestations of the biological activity of endotoxins but not its individual
components. Unlike protein toxins, they do not have organotropy and specificity of
action. Symptoms of intoxication in diseases caused by Gram-negative bacteria
are of the same type, and are associated with the action of the resulting
inflammatory mediators. The action of LPS is based on its non-specific lipid-
lipid and specific, due to the CD14 receptors, interaction with the membrane
components of different types of cells: platelets, granulocytes, red blood cells,
lymphocytes, monocytes and macrophages, which release biologically active
substances under the influence of LPS. LPS starts the synthesis of more than

15
20 different biologically active substances in a macroorganism, which
determine the pathogenesis of endotoxemia and have a pyrogenic effect.
The main point of its application are macrophages. The formation of large
doses of endotoxin is accompanied by inhibition of phagocytosis, symptoms of
severe toxicosis, weakness, shortness of breath, diarrhea, impaired cardiovascular-
system, decreased pressure, hypoglycemia, leukopenia, followed by leukocytosis,
platelet aggregation, hypothermia. With the formation of large amounts of
endotoxin in the blood due to increased destruction of a large number of Gram-
negative bacteria, the development of endotoxin shock is possible. With the
formation of small doses of endotoxin, mild toxicosis and an increase in body
temperature, stimulation of phagocytosis are noted. LPS refers to tymine-
independent antigens and causes polyclonal stimulation of B-lymphocytes,
activates the complement system in an alternative way, and is an adjuvant. Small
doses of endotoxin, which are constantly formed by representatives of the
normal microflora of the human body in the intestine, have a favorable
stimulating effect on the cells of the macroorganism’s immune system, which
leads to an increase in the nonspecific resistance of the macroorganism, its
resistance to infectious diseases, an increase in radioresistance and an increase
in the antitumor activity of cells. As a result of polyclonal stimulation and
activation of the complement system along an alternative pathway, the
macroorganism is in constant readiness to meet with a wide variety of microbes
and can resist them until specific defense factors are formed. On the other hand,
the prolonged presence of a polyclonal stimulant in a macroorganism can lead
to the inclusion of forbidden cell clones and the development of autoimmune
reactions. During the immune response, initially, 0-antibodies that do not have
antitoxic activity are formed on the administration of LPS. Symptoms of
intoxication decrease after the formation of antibodies to the R-core of the
polysaccharide and lipid A. Since they have the same structure in Gram-
negative bacteria, they try to use antibodies to treat septic processes caused by
these microbes. Unlike protein toxins, toxoids cannot be obtained from endotoxins.
Endotoxin causes a general intoxication of the body, showing inflammatory,
pyrogenic and allergenic action. The clinical picture that is caused by
endotoxins of different microorganisms is of the same type: the reaction of the
body is usually accompanied by common phenomena of intoxication such as
fever, headache, etc.
The close binding of endotoxin to the cells of the microorganism determines
its resistance to temperature and other external factors. To obtain endotoxin, it
is necessary to destroy the cell of the microorganism.
It should be remembered that a significant amount of toxin can lead to the
development of endotoxic shock and death of the patient, which occurs in the
mass death of microorganisms. Therefore, bactericidal antibiotics (penicillin for
typhoid) should not be used for diseases caused by Gram-negative microorganisms.

16
 The action of the toxin is determined on animals that are sensitive to this
toxin. For example, diphtheria toxin is tested on guinea pigs, botulinum toxin
on white mice, etc.
Features of infectious diseases are as follows:
• their etiological factor is a microbial agent;
Depending on the cause (on the etiological basis), infectious diseases are
divided into:
a) bacterioses, rickettsioses, mycoplasmoses;
b) viral infections;
c) mycoses.
Infestations (parasitic diseases) are divided into protozoa, helminth
infections and infections caused by arthropods.
• they are transmitted from the patient to healthy;
• induce immunity;
• have a number of common syndromes;
• characterized by cyclical course.
Infectious diseases are characterized by a cyclical course, which consists in
the presence of successively alternating periods based on the pathogenesis of
the disease. The duration of the periods depends both on the properties of the
microbe and on the resistance of the macroorganism, the characteristics of
immunogenesis. Even with the same disease in different individuals, the
duration of these periods can be different.
Clinical stages (periods) of infectious disease:
• incubation period (from lat. incubo - I rest or incubatio - without external
manifestations, hidden) is the period from the moment the infectious agent
penetrates the human body until the first precursors of the disease appear.
During the incubation period, the pathogen adapts to the internal environment
of the infected macroorganism and overcomes the protective mechanisms of the
latter. In addition to the adaptation of microbes, they multiply and accumulate
in the macroorganism, move and selectively accumulate in certain organs and
tissues (tissue and organ tropism), which are most susceptible to damage. On
the part of the macroorganism, already in the incubation period, its protective
forces are mobilized. There are still no signs of the disease in this period,
however, with special studies you can find the initial manifestations of the
pathological process in the form of characteristic morphological changes, metabolic
and immunological changes, the circulation of microbes and their antigens in
the blood. From the epidemiological point of view, it is important that a
macroorganism at the end of the incubation period can be an epidemiological
danger due to the release of microbes from it into the environment.
The duration of the incubation period has a certain duration, subject to
fluctuations both in the direction of decrease, and in the direction of increase. In
some infectious diseases, the duration of the incubation period is calculated in
hours, as, for example, with flu; with others – for weeks and even months, such

17
as for viral hepatitis B, rabies, slow viral infections. For most infectious
diseases, the incubation period is 1–3 weeks.
• The prodromal period (from Greek prodromes – harbinger) – the
manifestation of the first nonspecific symptoms of the disease, characteristic of
general intoxication of the macroorganism by the products of the vital activity
of microorganisms and the possible action of bacterial endotoxins released
during the death of the pathogen; they also stand out in the environment, the
patient in this period is already epidemiologically dangerous to others.
The prodromal period is not observed in all infectious diseases. It usually
lasts 1–2 days, but can be shortened to several hours or lengthened up to
5–10 days or more.
• The height of the disease – the manifestation of specific symptoms of the
disease. If there is a characteristic symptom complex in this period of the
development of the disease, the clinicians call this manifestation of the disease
a manifest infection, and in cases where the disease during this period proceeds
without pronounced symptoms, it is an asymptomatic infection. This period of
development of an infectious disease, as a rule, is accompanied by the release
of the pathogen from the body, as a result of which the patient poses an
epidemiological danger to others.
It was in this period that the specific pathogenic properties of microbes and
the response of the macroorganism find their fullest expression. This period is
often divided into three stages: 1) the stage of increase in clinical manifestations
(stadium incrementi); 2) the stage of maximum severity of clinical manifestations
(stadium fastigii); 3) the stage of attenuation of clinical manifestations (stadium
decrementi). The duration of this period varies significantly with different
infectious diseases, as well as with the same disease in different individuals
(from several hours to several days and even months). This period can end
fatally, or the disease goes to the next period, which is called the period of
extinction of the symptoms of the disease (early period of convalescence).
• The period of complete recovery (convalescence) – the cessation of the
pathogen reproduction in the patient’s body, the death of the pathogen and the
complete restoration of homeostasis. It is characterized by the absence of
clinical symptoms, the restoration of the structure and function of organs, the
cessation of the multiplication of the pathogen in the macroorganism and the
death of the microbe, or the process can go into carrier. The duration of the
convalescence period also varies widely even with the same disease and
depends on its form, severity, immunological features of the macroorganism,
and the effectiveness of the treatment.
There are: a) clinical recovery, in which only the visible clinical symptoms
of the disease disappear; b) microbiological recovery, accompanied by the
death the microbe; c) morphological recovery, accompanied by the restoration
of the morphological and physiological properties of the affected tissues and

18
organs. Usually, clinical and microbiological recovery do not coincide with the
complete restoration of morphological lesions that last a long time.
Sometimes, against the background of clinical recovery, carriage begins to
form – acute (up to 3 months), prolonged (up to 6 months), chronic (more than
6 months).
• Fatal outcome. It should be remembered that the corpses of infectious
patients are subject to mandatory disinfection, as they represent a certain
epidemiological danger due to the high content of microbial agent in them.
In the study of infection, there is also the concept of persistence (infection):
microorganisms enter the human organism and can exist in it without
manifesting themselves for a sufficiently long time.
For clinical purposes, infectious disease is usually divided by type. It is the
severity of the characteristics inherent in this nosological form. Typical forms
include cases of the disease in which there are all the leading clinical symptoms
and syndromes characteristic of the disease. Atypical forms include erased,
inapparent, as well as fulminant and abortive forms.
With erased forms, one or more characteristic symptoms are absent, and the
remaining symptoms are usually mild.
Inapparent (syn: subclinical, latent, asymptomatic) forms occur without
clinical symptoms. They are diagnosed using laboratory research methods,
usually in foci of infection.
Fulminant (from Lat. Fulminare - kill with lightning, or hypertoxic) forms
are characterized by a very severe course with the rapid development of all
clinical symptoms. In most cases, these forms end fatally.
In abortive forms, an infectious disease develops from the beginning
typically, but suddenly breaks off, which is typical, for example, of typhoid
fever in vaccinated people.
Biological method
Biological, or experimental, method of the diagnosis of infectious diseases
is the study of pathological material in laboratory animals.
The biological method of research is used when it is necessary:
1) to isolate pathogens that do not grow in nutrient media (viruses, rickettsiae);
2) to study some properties of isolated microorganisms (necrotic, toxigenic);
3) to isolate the pure culture of the pathogen from pathological material
contaminated with extraneous microflora, which inhibits the pathogen in
nutrient media. So, to isolate the pure culture of pneumococcus from sputum, in
which there are staphylococci, streptococci, it is emulsified in sterile broth and
administered intraperitoneally to white mice, whose body is very sensitive to
pneumococcus. Pneumococcus in the body of white mice multiplies very
quickly. In 6–8 hours it can be isolated from exudate (from Lat. Exsudo –
sweat, secrete; fluid leaking from small blood vessels into tissues and body

19
cavities in case of inflammation) of the abdominal cavity of a white mouse,
other microorganisms during this time do not have time to multiply.
Experimental contamination of laboratory animals is carried out for the
modeling of infectious disease, the study of the immune response of the
macroorganism, to study the effectiveness and safety of immunobiological and
medical products, to obtain immune diagnostic and therapeutic sera,
immunoglobulins.
Laboratory animals are also used as donors (from Lat. Dono – donate)
blood from which serum, plasma, erythrocytes, leukocytes are obtained. Blood
and its ingredients (from the Latin. Ingrediens - an integral part of the mixture)
are used for the production of nutrient media, the performing serological
reactions.
In microbiological practice white mice, rats, guinea pigs, rabbits are used
most often, chicken, pigeons, sheep, cats, dogs, hamsters, cattle, horses, pigs,
donkeys, monkeys are used rarely.
Recently, clean line of animals (inbred animals) have been used. They are
obtained by genetically close crossing. These are siblings for 20–40 generations,
selected on a specific basis. Genetic homogeneity of animals provides
uniformity of reactions of the body to the introduced pathogen.
In specialized laboratories, studies are conducted on non-microbial (sterile)
animals kept in aseptic conditions. The main purpose of the research is to study
the role of normal microflora in the physiological and pathological processes of
animals and humans. Science that studies the life of non-microbial macro-
organisms is called gnotobiology (from Greek. Gnotos - known, obvious and
biotos - life), and non-microbial macroorganisms - gnotobiots, or gnotobiotic.
The purpose, structure and equipment of the vivarium
Vivarium (from Lat. Vivus – alive), or experimental-biological clinic, is a
unit of the scientific institution in which the laboratory animals are kept and
sometimes bred for carrying out biomedical researches.
The vivarium is placed in a separate building or on the upper floors of the
laboratory building. In order to ensure anti-epidemic and anti-epizootic
regimes, the placement of vivarium must comply with the principle of division
into a clean and contaminated area.
The premises where they work with active drugs are potentially contaminated,
they are classified as contamination zone III. They must be completely isolated
from other rooms and the environment. The premises in which the infected
material is handled in sealed equipment, devices and protective boxes, are
qualified as conditionally contaminated and they belong to the zone II.
The premises of II and III zones are made especially tight, without windows
or with windows that are sealed and are not opened. The sealing these rooms
must be such that they can be treated with gaseous (vapor) chemicals. These
rooms should be provided by self-contained ventilation systems that dilute the

20
air, which provides the direction of air movement to these rooms, not the other
way around. The air that is vented by the ventilation systems from the premises
of zones II and III is filtered through a system of successively installed filters.
An autonomous wastewater disinfection system from these premises
ensures their high-temperature sterilization.
The buffer space is allocated to a separate group. These are the preparatory
ones serving the premises of zones II and III. They work with materials,
laboratory glassware and reagents, and also it is allowed the temporary
residence of the active material in a sealed package, which is intended for
transfer to the premises of zones II and III. These premises are considered
clean, they are referred to zone I.
In addition, there are general-purpose premises in the laboratory: lobbies,
locker room, office rooms, dining room, warehouses, maintenance room. This
group of premises is assigned to the 0 (zero) zone of the laboratory. At the
boundaries of the working areas of different zones barrier systems are created:
sanitary passes for people (rooms for the removal of clothing, air showers,
washing plant) and transfer facilities for the movement of materials, apparatus
and instruments for research (pass sterilizers for heat treatment, gas
transmission chambers for chemical treatment with a gas mixture) and transfer
baths ("diving chambers") for chemical treatment with disinfectant solutions.
They operate by technical systems from clean area premises.
The following departments are obligatory included in the vivarium:
quarantine, section for experimental animals, isolators, operating room,
manipulation, fodder kitchen, disinfection and washing department, warehouse
for clean spare equipment, sanitary unit (toilet, showers), domestic premises,
diagnostic room, service room, refrigerator.
The quarantine unit is intended for keeping animals purchased for the
vivarium. Animals are purchased from specialized nurseries. They are housed
in clean disinfected (pro-autoclaved) cages. The purchased animals must have a
passport indicating the date of departure and receipt of the animals, the cage
number, quantity, age and body weight of the animal, the date and results of the
clinical examination. If the animals are purchased from a specialized nursery
located in the same town, they are kept in that compartment for 3 days to adapt
to new conditions. If the animals are purchased from nurseries located in other
cities, or non-specialized nurseries, the quarantine period is set according to the
incubation period for the most common diseases for these animals. Quarantine
term: for mice and rats – 17 days, cats and dogs - 30 days, guinea pigs, rabbits,
birds and other animals – 21 days.
During the quarantine, animals are monitored daily for clinical supervision.
The quarantine premises are cleaned and disinfected after each batch of animals
transferred for work and after each case of detection of infectious disease. At the
end of the quarantine period, animals are transferred to the experimental sections.

21
 Isolators are designed to contain animals with suspected infectious diseases
or ill animals, the destruction of which is undesirable under the conditions of
experience.
Constructive features of operating and manipulation are determined in each
case depending on the task and purpose of scientific research.
The fodder kitchen consists of two adjacent premises for processing and
production of animal feed.
The disinfection and washing compartment are two adjacent rooms
connected by a passing autoclave or a passing dry chamber. It is intended for
sterilization of inventory, litter, cages after mechanical cleaning.
Diagnostic and service rooms are laboratory facilities for research on the
quality control of animals, feed, and for maintaining documentation.
The refrigeration chamber is intended for storage of carcasses of animals for
pathological anatomical examination.
In rooms where animals are kept or experiments are carried out, as well as
in the fodder kitchen and in the disinfection and cleaning department, the floor
is made of waterproof material, without plinths with a slope to the openings
connected to the sewer. The walls from floor to ceiling are covered with glossy
tile, the ceiling is painted with oil paint. The doors have a smooth surface, painted
with oil paint. The upper part of the door is glazed. Lighting and microclimate
(temperature, ventilation) in the premises must meet the established standards.
Rules for the keeping and care of laboratory animals
The correct keeping of laboratory animals and the proper nutrition of
animals is of great importance for obtaining reliable results of the experiment.
Disruption of the mode of retention and diet causes the weakening of the body
of animals, increasing their sensitivity to pathogenic microorganisms, which
leads to distortion of the results of the study and incorrect findings. Cages with
laboratory animals, boxes, enclosures are placed in the area provided by the
established standards.
All work on the care and maintenance of laboratory animals is carried out in
accordance with the approved regulations, which determine the time for sanitary
treatment of premises and equipment, feeding, carrying out experimental works
and manipulations. Animal care and hygiene personnel should wear work
clothes (trousers or overalls with elastic seals at wrists and ankles, headgear
should cover hair completely, gloves, special footwear), goggles and respirators.
Personnel serving animals infected with pathogens of particularly dangerous
infections should wear a bathrobe, rubber apron, sleeves, rubber gloves, rubber
boots or galoshes on top of their clothing, and goggles and a cotton gauze mask
if necessary.
Cages and equipment are cleaned and washed daily after pre-decontamination.
Contaminated litter and other waste from cages are collected in special metal

22
tanks with lids. The tanks are tightly closed and transferred to the washroom
and disinfection room. The waste is disinfected or incinerated. The methods of
disinfection, disinsection and autoclaving are set on a case-by-case basis depending
on the animal's condition (healthy or infected) and the type of infection.
Laboratory animals are fed 2–3 times a day according to the standards
developed by the research institutions. The daily diet is made so that different
types of feed are given alternately. The main food for laboratory animals is
cereal grains (oats, wheat, corn, millet), oilseeds (sunflower, flax, hemp) and
legumes (peas). They are given natural or crushed. It is sterilized in an
autoclave or in a dry oven before being fed to animals.
Green, juicy feed, milk, fat, vitamins, and mineral salts are also required for
good nutrition. There should always be drinking water in the cages of the animals.
All actions related to the care, keeping animals, carrying out medical and
biological experiments (surgery, blood collection, necessary killing of animals) are
performed in accordance with the rules of humane treatment for laboratory animals.
Preparation of laboratory animals for experiment
For the experiment, a group of animals of a certain species, age, body
weight is selected, sometimes, under the conditions of the experiment, one sex.
All animals must be healthy. They are placed in clean disinfected cages and
weighed for several days before the experiment, thermometry and marking of
animals are carried out.
Weighing. For weighing mice, rats, guinea pigs dial scales are used
(without weights) with a loading capacity up to 1 kg; for weighing rabbits cup
scales with tare wooden boxes or cardboard boxes (with known weight) with a
capacity up to 5 kg are used.
Thermometry. Thermometry is performed with a medical mercury thermometer
(there is a special small triangular thermometer for mice) in the rectum. Normal
temperature in different animals is not the same: in white mice – 37–39 °C,
white rats – 38,5–39,5 °C, guinea pigs – 37,3–39,5 C, rabbits – 37,7–38,0 °C.
Fixation of laboratory animals. During the biomedical experiment, the
animals are fixed (limiting their mobility) in a posture that makes it convenient
to perform the intended manipulation. There are several ways of fixing animals.
Fixation of animal (rabbit, guinea pig) with a towel. The animal is tightly
wrapped in a towel. In this case, the front paws are pressed to the chest, the
back - to the abdomen, the head remains free.
Fixation of the head of a rabbit. To do this, use a box (wooden box with a
movable lid and the top of the front wall). In the front wall cut a hole with a
diameter of 5 cm so that it was located in the lower stationary part of the wall
and in the upper movable. The rabbit is put in a box, and his head is brought out
through this opening. Moving parts of the box fix the trunk and head of the rabbit.
Fixation of animals (rabbit, guinea pig, rat) in a supine or abdominal
position. To do this, use a special machine. This is a normal board on low legs,

23
the size of which corresponds to the size of the animal. The side edges of the
board have two holes or two hooks to secure the loops (fixation of the legs). At
the narrow edge of the board there is a device for fixing the head.
Fixation of guinea pig with hands. The manipulation is performed together.
The assistant fixes the animal, and the experimenter performs the appropriate
manipulation. The guinea pig is taken with the left hand so that the second
finger of the left hand is under the neck, and the first and third fingers – under
the forelimbs of the animal, and rotate it with the belly up or out. With the right
hand, the pig is stroked to the abdomen until it calms down. The hind legs of
the animal are held by the right hand.
Fixation of rats. The fold of the skin on the nape of the neck is taken with a
cornice and pressed tightly the animal's head against the table surface. They
take the rat's tail with the other hand and, barely lifting it above the table surface,
hold it in such a position that the head is slightly drawn away by the cornice.
Fixation of mice. Mouse is let on the surface of the table, holding it by the
tail with the first and second fingers of the right hand. After straightening the
tail with a quick movement of the left hand, it is grabbed by the fold of skin out
of her ears so that it cannot turn her head. Lifting the mouse over the table, the
assistant holds it with one hand by the tail and the other by folding the skin on
the nape in a comfortable position for the experimenter.
It is possible to work with mice without the assistant, fixing them with the
left hand: the first and second fingers of the left hand hold the animal by the
fold of skin on the nape of the neck, and the other three fingers press the tail
and the skin of the back with the wrist. With this method of fixation, the right
arm remains free, allowing various manipulations to be performed.
Methods of infection of laboratory animals
Depending on the purpose of the study, different methods of infection are
used: intradermal, cutaneous, subcutaneous, intramuscular, intraperitoneal,
intravenous, oral, intranasal, intracerebral, infestation in the eye, intracardiac.
In all ways of infection a syringe is used. The area of the body on which the
injection or incision is made is called the operative field. When the material is
introduced through the skin, the surgical field is pre-released from wool (cut,
shaved, plucked or treated with an epilator – a mixture that causes hair to fall
out). Before injection, the operating field is treated with a 70 % solution of
ethyl alcohol or alcohol solution of iodine. To prevent the formation of
contaminated aerosol when removing air from the syringe, a cotton swab
moistened with 70 % ethyl alcohol solution is put on the needle. The
contaminated cotton swab is lowered into a jar of disinfectant solution. (When
working with infectious agents, it is forbidden to remove air from the syringe,
so fill the needle with water from the sterilizer).

24
 Intradermal method of infection. For this thin sharp needles with a small
bevel are used. Before inserting the material, the skin is stretched with the first
and second fingers of the left hand, and the needle of the syringe is inserted by
the right hand at a very acute angle. In this case, the end of the needle shines
through the epidermis, and a hump is formed at the site of the introduced
material; the skin on its surface is transparent and porous, so it is compared to a
lemon peel. The material is injected into the skin of the back or abdomen in a
volume of 0.1 ml.
In this way they put dermonecrotic and allergy tests.
Cutaneous infection method. After removal of wool, more often in the
abdomen, the skin is scarified (from Lat. Sagifis – scratch) with a scalpel or
Jenner pen. The material is applied to the skin and rubbed into scratches. The
method is used to identify pathologic agents of tularemia, plague.
Subcutaneous method of infection. The skin at the site of material insertion
is lifted with the first and second fingers of the left hand (Fig. 3). The needle of
the syringe is inserted at the bottom of the formed folds of the skin. Puncture
the skin, insert the needle a few millimeters, then slightly turn it right or left
and slowly enter the material. The direction of the needle is changed so that the
material does not leak through the puncture. The skin fold is lowered, a cotton
wool moistened with alcohol is placed at the injection site, the needle is quickly
removed.

Fig. 3. Subcutaneous method of infection of laboratory animals

The material is introduced to the rabbits and guinea pigs under the skin of
the back or lateral surface; rats, mice - under the skin of the back, waist, nape.
Rabbits are injected with up to 30 ml of fluid, guinea pigs up to 15 ml, rats up
to 10 ml, mice up to 1 ml.
Intramuscular method of infection (Fig. 4). The site of insertion of the
material is the outer upper third of the hind paw (the area of the body with the
most developed muscles). The muscle fold is taken with the first and second

25
fingers of the left hand, and the needle of the syringe is injected almost at right
angle deep into the muscles. Rabbits are administered up to 8 ml of fluid,
guinea pigs – 5 ml, rats – 3 ml, mice – 0.5 ml.

Fig. 4. Intramuscular method of infecting laboratory animals

Intraperitoneal method of infection (Fig. 5). The helper holds the animal head
down, with the intestines moving to the side of the diaphragm, reducing the
possibility of their damage. The material is introduced into the lower third of the
abdomen to the left of the midline. In this case, the needle of the syringe is kept
perpendicular to the abdominal wall, injected until a feeling of "failure", then
remouve the needle upright and inject the material into the abdominal cavity:
rabbits – up to 30 ml, guinea pigs – up to 10 ml, rats – up to 5 ml, mice – up to 2 ml.

Fig. 5. Intraperitoneal method infection of laboratory animals

This method is used to detect the exotoxin of the botulinum pathogen.
Intravenous method of infection. Rabbits are infected in the marginal vein of
the ear. After wool is removed, the ear is clamped at its base to cause vascular

26
flushing. The syringe needle is inserted at an acute angle. If the needle enters
the vein, the material is easily inserted. Rabbits are injected with up to 20 ml of fluid.
Rats and mice are infected in the lateral vein of the tail. Before introducing
the material, the tail of the animals is immersed in water heated to 50 °C; the
veins are swollen and shine through the skin. The syringe needle is inserted at a
very acute angle. Rats were injected with up to 6 ml of fluid, mice were
injected with up to 0.5 ml.
The force of action of microbial exotoxins is determined in this way (DLM, DCL).
Oral infection is infection through the digestive tract. There are two ways to
infect animals through the mouth. The material intended for infection is added
to the food or to the drinking liquid (milk). This method is simple and
convenient, but does not allow to take into account the amount of material that
gets into the animal's body. Therefore, the material is more often injected with a
syringe. To do this, take a needle with a thickening on the end in the form of
olive. It is more convenient and safer to use an elastic probe. Its outer end is
attached to the syringe, and the lower end is inserted into the lower esophagus
or directly into the stomach. In this case, rats and mice are kept upright, and
rabbits and guinea pigs are wrapped in a towel and sit on the assistant's lap. In
this way, rabbits are injected with 3.5–5 ml of fluid, guinea pigs 2.5–3.5 ml,
rats up to 3.5 ml, mice 1 ml.
This is how the effects of enterotoxins of pathogens of food poisning are studied.
Intranasal method of infection is infection through the respiratory tract. The
animal is fixed on a preparation board. A cotton swab moistened with ether or
thiopental is broucht to the nose. After signs of light anesthesia appear in the
animal, the needle of a syringe or pipette is introduced into the nostrils of the
mice to a depth of 1–1.5 mm, rats – 2–3 mm, rabbits and guinea pigs – 4 mm.
The material is injected with small drops. To avoid damaging the mucous
membranes, blunt needles schould be used.
In this way, the influenza virus is detected in the material being examined.
Intracerebral (subdural) method of infection. The material is injected through
the crown of suckling mice under the dura. Intracerebral infection is also carried
out to rabbits, guinea pigs, mice. At the same time, after opening the skin in the
middle of the forehead, pierce the bone with Frank's needle, then inject the needle
of the syringe into this hole. Rabbits are injected with 0.4 ml, guinea pigs 0.2 ml
of infected fluid. Mice are pierced with a needle from a syringe and injected with
0.05 ml of infected fluid.
In this way, viruses of encephalitis, and polio, Coxsackie, ECHO viruses are
detected in the pathological material.
It is possible to incorporate infected material through the eye in various ways.
The lower outer corner of the eyelid of the animal is drawn with tweezers (in the left
hand), with the right hand (from a pasteur pipette 1–2 drops or a bacteriological

27
loop) the material is applied to the conjunctiva and distributed on the conjunctiva,
moving the eyelids with tweezers. In the subconjunctival method, the material is
injected with a fine needle under the conjunctiva (0.1–0.2 ml). You can also insert the
material into the scarified cornea of the eye by rubbing it with a glass spatula.
The method is used for performing a keratoconjunctival test in the diagnosis
of shigellosis, colienteritis caused by enteroinvasive Escherichia.
Intracardiac infection. The material is injected into the ventricle of the heart.
This method is used to reproduce anaphylactic shock.
All methods of animal contamination should strictly adhere to the
methodological guidelines governing the preparation, conduct of the experiment,
the decontamination of pathological material and tools, and the rules of conduct
of the staff in order to prevent intra-laboratory contamination.
Methods of blood collection in laboratory animals.
Treatment and selection of blood components
Manipulations (from Latin. Manus - hand, action of hands at performance of
work) at blood sampling from animals are different, they depend on a kind of
animals and methods of blood sampling. A small amount of blood is taken from
the veins of the ear, tail, inner surface of the wing. More blood is taken from the
heart of a rat, a rabbit, a guinea pig, an elbow vein (in a monkey), a thigh vein, a
carotid vein of a horse, cattle. In special cases, conduct a total (from Lat. Totalis –
whole) bleeding. The disadvantage of this method is that the animal dies.
Before blood is collected from laboratory animals, on the area of the body
where the puncture is to be punctured the wool is first removed, degreased and
disinfected with alcohol, ether or alcohol iodine tincture. After taking the blood, a
cotton swab impregnated with an alcoholic iodine solution is applied to the
injection site, and the animal is injected subcutaneously with a sterile isotonic
sodium chloride solution or glucose solution heated to 37 °C in a volume that is
2 times greater than the volume of taken blood.
Blood collection from the heart. The rabbit, rat, guinea pig are fixed on a
special table with the belly up. With the second finger of the left hand feel the
heart impulse and in this place enter the needle of the syringe. If the needle hits
the heart, blood immediately begins to fill the syringe. The amount of blood that
can be taken depends on the species of animal. The rabbit can take 25–30 ml, the
guinea pig – 10–12 ml, the rat – 6–8 ml. To prevent blood clotting, the syringe
and needle are rinsed with sterile sodium citrate before blood collection, and the
manipulation is performed as soon as possible.
Blood collection from a vein. In a rabbit blood is taken from the marginal vein
of the ear. To do this, it is fixed with a towel and kneel down. They remove wool
on the outer surface of the ear, disinfect the skin, clamp a vein at the root of the
ear, causing blood flow to be delayed and a vein swollen. The needle is inserted

28
into the vein without a syringe, and the vein is released. Blood dripping from the
needle is collected into a sterile test tube.
In white mice and white rats, blood is taken from the tail vein. Animals are
fixed in metal mesh of appropriate size. The tail is first heated in water at a
temperature of 45–50 °C, then amputated (from the Lat. Amputatio – cut off) the
tip of the tail. Blood flowing from the stump is collected into a sterile test tube.
To stop the bleeding the stump is treated with hydrogen peroxide.
Large animals (horses, rams, bulls) are first anesthetized (from Greek.
Narcosis - numbness), and then blood is taken from the carotid artery.
Total revelation. The animal is fixed with the belly up, give the animal
anesthesia. After that, they make a section along the middle line of the neck, dilute
the edges of the wound, then separate the muscles, expose the carotid artery and
impose on it 2 ligatures (from the Latin Ligatura - ligament). Below the second
ligature impose clamp. Between the ligature and the clamp insert a cannula (glass
tube with the drawn capillary). A rubber tube is placed on the extended part of the
cannula and the end of the cannula is lowered into the vial. The clip is removed, the
blood flows freely into the vial. With total bleeding, the rabbit you can take
120–150 ml of blood, the guinea pig – 30–40 ml, the rat – 20–25 ml.
In laboratory practice, both whole blood and its individual constituents are
widely used: erythrocytes, serum, and plasma.
Production of defibrinated blood. Animal blood is collected into a sterile
glass bead vial. The vial is shaken within 10–15 minutes, causing the fibrin to
settle on the surface of the necklace and the blood lose its ability to coagulate.
Defibrinated blood is poured into another sterile vial. It is used for the
manufacture of complex nutrient media, as well as for obtaining a suspension
of red blood cells.
Production of suspension of erythrocytes. The defibrinated blood is poured
into a centrifuge tube, closed with a stopper and centrifuged at 2 000 –3 000 rpm
for 10–15 minutes. The erythrocytes settle to the bottom of the tube, and the
transparent part of the blood forms a supernatant. After centrifugation, the liquid
level in the tube is indicated by a pencil, the liquid portion is aspirated with a
Pasteur pipette with a rubber pear, and a sterile isotonic sodium chloride solution
is poured into the test tube and centrifuged again. After centrifugation, the
supernatant is aspirated and poured into a disinfectant solution, and a sterile
isotonic solution is poured again. Washing the erythrocytes with sterile isotonic
solution is repeated 3–4 times. The last dose of wash fluid should be colorless.
3–5 % suspension is prepared from the washed erythrocytes, which can be stored
in the refrigerator at 3–4 °C for 5–6 days.
The erythrocyte suspension is used to performe reactions to detect antibodies in
the patient's serum, to detect viruses in pathological material.

29
 Production of blood serum. In order to obtain a clear serum, the animals
should be fasted. Lipimia (accumulation of excess fat) occurs after ingestion of
the animal feed and sometimes with repeated bloodletting at short intervals.
Serum obtained from such blood will be cloudy (chylous), so the results of
serological reactions may be incorrect. The blood is collected into a test tube,
preventing the formation of foam. For this purpose, a stream of blood flowing
from the blood vessel is directed to the test tube wall. The tube with the blood can
be left at room temperature for 3–4 hours. For more efficient and accelerated
coagulation, the blood is placed in a thermostat at 37 °C for 20–30 minutes. (It is
not possible to leave blood in the thermostat longer because hemolysis will
occur). The formed blood clot is separated from the test tube walls with a sterile
bacteriological loop and placed in the refrigerator for 1 hour or centrifuged. Well-
settled serum is obtained by settling blood for 18–20 hours. Serum above the
precipitate can be stored for no longer than 48 hours (with longer storage
hemolysis occurs). If the serum is used later than 48 hours, it is transferred to a
sterile tube with a sterile pipette.
Production of citrate blood. Pour 1 ml of 5 % sodium citrate solution into a
blood collection tube and select 10 ml of blood. The tube is closed with a stopper
and the tube contents is mixed. Citrate blood does not clot.
Production of blood plasma (from the Greek. Plasma – molding, design).
Plasma is made from citrate blood. For this purpose, citrate blood is placed in the
refrigerator for 18–20 hours or centrifuged. A layer of slightly cloudy fluid of
light yellow color is formed above the precipitate of the formed elements of blood
(erythrocytes, platelets, leukocytes). Plasma differs from serum in that it contains
fibrinogen protein. Plasma is used in laboratory practice to detect the plasma-
coagulating properties of staphylococcus.
Autopsy of carcasses of laboratory animals,
maintenance of the protocol of autopsy
The dissection of corpses of infected laboratory animals is done to determine
the cause of death of the animals, isolation of the introduced pathogen, identify
the mechanism of spread of germs in the body and the location of their
localization, study of pathoanatomical changes resulting from the infection. In the
case of an experimental infection, the opening the animal is performed after its
death. But there are cases when an animal is killed. There are several ways to do
this. The simplest is that the animal is placed in a container, balls of cotton wool
soaked in ether or chloroform are lowered into it, and tightly closed.
The animal dies in 3–5 minutes. To kill rabbits an air embolism is used (from
the Greek embole - insertion, occlusion of blood or lymphatic vessels by foreign
particles – emboli brought by blood or lymph, which leads to impaired blood
supply to organs and tissues); sometimes mice are decapitated (from Lat.

30
Deremoval - destruction and caput - head). The dissection of the corpse is carried
out immediately after the death of the animal, to prevent the penetration of germs
from the gut into the blood and other organs. In the absence of such possibility,
the corpses are stored in the refrigerator for no more than 1 day.
Experimental study of infected animals with autopsy is performed in the
following sequence:
– preparation of the animal for opening;
– fixation of the corpse;
– inspection of external coverings;
– opening and examination of the chest cavity;
– conducting a study of the peritoneal cavity;
– preparing smears-imprints;
– selection of pathological material for seeding on nutrient media.
According to the stages of autopsy, the state of the outer coverings of the
body, subcutaneous fat, lymph nodes; presence of exudate in the cavities of the
pericardium and pleural (pericarp), appearance of the chest (color, size,
consistency of the heart and lungs), the presence of inflammation, hemorrhage; in
the abdomen – the presence of exudate, the appearance of the liver, spleen,
adrenal glands, mesenteric lymph nodes are noted in the protocol.
Algorithm "Preparation of an animal corpse for opening":
Warning! Always observe the safety precautions when working with
laboratory animals!
put on rubber gloves;
place the animal in a glass or metal jar;
moisten a cotton swab with ether or chloroform and place it in a jar with the
animal;
close the jar with a lid without holes or a towel, hold for 3–5 minutes;
remove the animal's corpse from the jar with long tweezers, place it on a
preparation board or tray filled with paraffin.
Algorithm "Carrying out the opening of an animal's corpse" (Fig. 6):
cover the surface of the preparation board with a gauze soaked in 3 %
chloramine solution;
put the body of the mouse on the gauze belly up;
fix the mouse pads with preparatory needles;
take a cotton ball with tweezers, moisten it with 3% chloramine solution,
wipe the surface of the animal body;
tighten the skin with surgical forceps;
cut the skin with scissors in a straight line from the pubis to the mandible
and to each paw;
Separate (from the Lat. separatio - separation) pieces of skin, pulling them
with surgical forceps and clipping with a scalpel;

31
 Warning! When separating the skin, it is necessary to expose the anterior
and lateral walls of the chest and the walls of the abdominal cavity; inguinal
lymph nodes are separated along with the skin, the submandibular and cervical
remains on the muscles of the neck.
wipe the tools with a cotton ball soaked in water, place the ball in a jar with
disinfectant solution;
lower the instruments in a jar with alcohol, remove them and burn them in
the flame of the alcohol;
make two mutually perpendicular cuts of the peritoneum with scissors and
open the abdominal cavity;
cut the pectoral muscles, ribs on both sides between the bone and cartilage
with scissors, cut the sternum and drop the muscle and bone piece on the
animal's neck;
wipe the instruments with water-soaked cotton wool, put them in a jar
with alcohol.

Fig. 6. Stages of dissection of an animal:

1 – skin incision; 2, 3 – section of abdominal cavity, 4 – section of the chest cavity

32
 Warning! In parallel with the opening the corpse of the animals keep a
protocol of the opening. The protocol indicates the age of the animal, the
number, time and place of infection, the material that was infected, the time of
death of the animal (or time of killing).
Algorithm "Selection of section material, preparation of smears-imprints,
seeding section material on a nutrient medium":
burn anatomic forceps and scalpel in an alcohol flame;
Warning! Do not use surgical forceps, it destroys tissues!
take the organ with tweezers, cut a piece with a scalpel;
make smears-imprints of organ pieces on one slide in the following order:
lymph nodes (4 prints), lungs (3 prints), spleen (2 prints), liver (1 print);
lower a piece of lungs into a glass of water (a piece of healthy lungs floats
on the surface of the water; a piece of patient lungs sinks);
light the top of the heart with a hot scalpel;
pierce it with a sterile Pasteur pipette, collect blood into the pipette;
apply 1 drop of blood to a slide;
seed 1-2 drops of blood into a test tube with sugar broth;
lower the pipette into a disinfectant solution (5% carbolic acid solution);
make a blood smear from the drop applied to the slide;
lower the slide with smear-imprints and a smear of blood into the
Nikiforov solution for fixation;
remove the slides after 20 minutes, air dry;
stain smear-imprints with methylene blue;
stain smear of blood after Romanovsky;
lower the tools into a disinfectant solution;
remove the preparation needles, lower them into the disinfectant solution;
remove the animal's body with a gauze with tweezers, lower it into a
disinfectant solution;
pour over the surface of the paraffin with alcohol and set it on fire;
clean up the workplace: hand tools and a tank with an animal corpse to an
autoclave technician (the corpse can be burned)
wipe with a disinfectant solution the table surface;
remove the rubber gloves by twisting them, lowering them into a
disinfectant solution;
put the Petri dishes wit culture in the thermostat;
Examine the smears and blood smears under the microscope.
Specific goals:
To apply biological research method for isolation from pathological
material of infectious disease agent and to identify pathogenicity factors with
registration of the research result.

33
 Student should be able:
1. Prepare a laboratory animal for the experiment: mark, weigh, fix.
2. To conduct thermometry.
3. To infect, to open the carcasses of laboratory animals, to keep a protocol
of the opening.
4. To make smears-imprints.
5. Selection and seeding section material on nutrient media.
6. To defuse the carcasses of animals after opening.
Theoretical questions:
1. What is a biological method for the diagnosis of infectious diseases.
2. Methods of infection of laboratory animals.
3. Methods of taking blood from a laboratory animal and obtaining from it
defibrinated blood, citrate blood, serum, plasma.
Practical tasks performed in the class:
1. Preparation of the workplace.
2. Carrying out fixing, marking, weighing of laboratory animals.
3. Preparation of the animal (white mouse) for opening, carrying out the
opening of the corpse, drawing up the protocol of the opening.
4. Preparation of smear-imprints.
5. Selection of section material for microbiological research, seeding it on
nutrient media.
6. Carrying out disinfection of corpses after opening.
7. Registration of the protocol.
Short guidelines for working on a practical lesson:
At the beginning of the class, the level of preparation of students for the
class is checked. Students study the structure and equipment of the vivarium,
the sanitary and hygienic requirements for the keeping laboratory animals; rules
for the care of laboratory animals, preparation of laboratory animals for
experiment, methods of fixation of animals and methods of their infection,
methods of blood collection in laboratory animals, treatment and separation of
blood components.
At the end of the class, test control and analysis of the results of each
student's independent work are conducted.
Targeted learning objectives:
1. The causes of the pathogenic properties of conditionally pathogenic bacteria are:
A. Biochemical properties of the strain.
B. Toxins of microorganisms.
C. The complex of properties of microorganisms and features of the human body.
D. Decrease in immunity of the macroorganism.
E. Adhesive properties of microbial cells.

34
2. Patient S. has acute gonorrhea. it is known from the anamnesis that the
patient had previously suffered from gonorrhea and was completely cured. To
which group of infections can a new disease be attributed?
A. Autoinfection. C. Reinfection. E. Mix-infection.
B. Relapse. D. Superinfection.
3. The factors of spread of pathogenic microorganisms in the macroorganism are:
A. Neuraminidase, hyaluronidase. D. Exo- and endotoxin.
B. Capsule substance and coagulase. E. Spore.
C. Flagella.
4. Patient B. is registered at the skin and venereological dispensary where he
undergoes treatment for syphilis, re-infected with T. pallidum. Which group of
infections can be attributed to this new disease?
A. Secondary C. Reinfection E. Mix-infection
B. Superinfection D. Relapse
5. To produce serum test tubes, you can:
A. Leave at room tº for 3–4 hours.
B. Leave in thermostat at tº = 37 ºС for 20–30 minutes.
C. Leave in thermostat at tº = 37 ºC for 2–3 hours.
D. Leave in the refrigerator for 12 hours.
E. Correctly 1.2.
6. Bacterial endotoxins are generally:
A. Plasma-soluble proteins D. Cell membrane components
B. Cell wall components E. Nucleoid components
C. Coded for by a temperate phage
7. The role of bacterial capsules as virulence factors is usually related to their
ability to interfere with:
A. Antibody binding.
B. B lymphocyte activation.
C. Antibacterial penetration of bacterial cells.
D. Phagocytosis.
E. The release of interferon-γ and other macrophage-activating cytokines.
8. Which of the following bacterial enzymes attacks the intercellular cementing
materials of human cells?
A. Myeloperoxidase C. Hemolysins E. Catalase
B. Streptokinase D. Hyaluronidase
9. Septicaemia is:
A. Pus in blood. D. Presence of viruses in blood.
B. Toxin in blood. E. Multiplication of bacteria in blood.
C. Presence of bacteria in blood.
10. Lowering virulence of bacteria is known as:
A. Pathogenicity. C. Attenuation.
B. Exaltation. D. Toxigenicity. E. Communicability.

35
11. Vertical transmission of bacteria means:
A. Transplacental transmission. D. Transfusion transmission.
B. Man to man transmission. E. Inhalation.
C. Animal to man transmission.
12. Endotoxin of Gram negative bacteria has all, EXCEPT:
A. Carbohydrate. C. Lipid. E. All of the above.
B. Protein. D. Inorganic substance.
13. All of the following are characteristics of endotoxins, EXCEPT:
A. Heat stable. D. Produced by Gram negative bacteria.
B. Action often enzymatic. E. Protein-polysaccharide complex.
C. Can not be toxoided.
14. The patient was taken to hospital with purulent wound infection. S.aureus
and P. aeruginosa were isolated from wound. What type of infection can this
disease be attributed to?
A. Reinfection. C. Relapse. E. Mixed infection.
B. Secondary infection. D. Superinfection.
15. Relapsing fever caused by Borrelia causasica is found only in certain
territories where the carrier is a kind of mite Alectorobius. What is such
infection called?
A. Mixed infection. C. Secondary infection. E. Reinfection.
B. Endemic. D. Superinfection.
16. The patient as a result of the activation of own microflora in the oral
mucosa, there developed purulent inflammation of periodontal tissues. What
type of infection can this disease be attributed to?
A. Autoinfection. C. Endemic.
B. Superinfection. D. Mixed infection. E. Secondary infection.
REFERENCES

1. Tsyganenko A.Ya., Vasilchenko V.N., Kovalenko N.I. Practical

exercises of morphology of bacteria. Short textbook for students of high
medical schools. – Kharkiv: KSMU, 2012. – 270 p.
2. USML Step 1. Lecture notes. – New York, 2018. – 2568 p.
3. Murray P.R. Manual of Clinical Microbiology. 8th Ed / P. R. Murray,
K. S. Rosenthal, M. A. Pfaller. – Washington : ASM Press, 2015. – 848 p.
4. Ananthanarayan. Textbook of Microbiology. / Ananthanarayan,
Paniker. – 9th Ed – Orient Blackswan, 2013. – 657 p.
5. Manual of Clinical Microbiology / Karen C. Carroll et al. – 10th
Edition. – Vol. 2. – Washington, DC : ASM Press, 2011. – 2630 p.
6. Manual of Clinical Microbiology / Karen C. Carroll et al. – 10th
Edition. – Vol. 2. – Washington, DC : ASM Press, 2011. – 2630 p.

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 Навчальне видання

ІНФЕКЦІЯ.
ІНФЕКЦІЙНИЙ ПРОЦЕС
Методичні вказівки з дисципліни
"Мікробіологія, вірусологія та імунологія"
для студентів II і III курсів медичного
та стоматологічного факультетів
з англійською мовою викладання
Упорядники Коваленко Наталія Іллівна
Замазій Тетяна Миколаївна

Відповідальний за випуск Н. І. Коваленко

Комп'ютерний набір Т. М. амазій

Комп'ютерна верстка О. Ю. Лавриненко

Формат А 5. Ум. друк. арк. 2,3. Зам. № 19-33854

______________________________________________________________
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Свідоцтво про внесення суб’єкта видавничої справи до Державного реєстру видавництв,
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